公开(公告)号：US20150337295A1

公开(公告)日：2015-11-26

申请号：US14708048

申请日：2015-05-08

Applicant: FLUIDIGM CORPORATION

Inventor： JASON A.A. WEST ,  BRIAN FOWLER ,  TZE-HOWE CHARN ,  CHRISTIAN F. JOHNSON ,  MARC A. UNGER

IPC: C12N15/10 ,  C12Q1/68

CPC classification number: C12Q1/6855 ,  C12N15/1065 ,  C12Q1/6806 ,  C12Q2565/501 ,  C12Q2521/301 ,  C12Q2521/501 ,  C12Q2525/155 ,  C12Q2537/162 ,  C12Q2565/543 ,  C12Q2535/122

Abstract: This disclosure provides a method of forming tagged nucleic acid sequences. A target polynucleotide is immobilized on a solid support; a recognition-oligonucleotide is hybridized thereto; the recognition-oligonucleotide-target polynucleotide hybrid is cleaved; and an adapter nucleic acid is ligated to the cleaved target polynucleotide, thereby forming a tagged nucleic acid sequence. Also provided is a method of forming a tagged single stranded cDNA; a method of forming a plurality of tagged heterogeneous nucleic acid sequences; a library of recognition-oligonucleotides; and methods for amplifying a cDNA sequence immobilized on a solid support. These methods and products can be used alone or in combination for integrated single cell sequencing, and can be adapted for use in a microfluidic apparatus or device.

Abstract translation: 本公开提供了形成标记的核酸序列的方法。 靶多核苷酸固定在固体支持物上; 识别寡核苷酸与其杂交; 识别寡核苷酸靶多核苷酸杂交体被切割; 并将衔接子核酸连接到切割的靶多核苷酸上，从而形成标记的核酸序列。 还提供了形成标记的单链cDNA的方法; 形成多个标记的异源核酸序列的方法; 识别寡核苷酸文库; 以及用于扩增固定在固体支持物上的cDNA序列的方法。 这些方法和产品可以单独使用或组合使用以用于集成的单细胞测序，并且可以适用于微流体装置或装置。